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Jodar M.,Wayne State University | Selvaraju S.,Wayne State University | Selvaraju S.,Indian National Institute of Animal Nutrition and Physiology | Sendler E.,Wayne State University | And 2 more authors.
Human Reproduction Update | Year: 2013

background: Spermatozoa are highly differentiated, transcriptionally inert cells characterized by a compact nucleus with minimal cytoplasm. Nevertheless they contain a suite of unique RNAs that are delivered to oocyte upon fertilization. They are likely integrated as part of many different processes including genome recognition, consolidation-confrontation, early embryonic development and epigenetic transgenerational inherence. Spermatozoal RNAs also provide a window into the developmental history of each sperm thereby providing biomarkers of fertility and pregnancy outcome which are being intensely studied. methods: Literature searches were performed to review the majority of spermatozoal RNA studies that described potential functions and clinical applications with emphasis on Next-Generation Sequencing. Human, mouse, bovine and stallion were compared as their distribution and composition of spermatozoal RNAs, using these techniques, have been described. results: Comparisons highlighted the complexity of the population of spermatozoal RNAs that comprises rRNA,mRNAand both large and small non-coding RNAs. RNA-seq analysis has revealed that only a fraction of the larger RNAs retain their structure. While rRNAs are the most abundant and are highly fragmented, ensuring a translationally quiescent state, other RNAs including some mRNAs retain their functional potential, thereby increasing the opportunity for regulatory interactions. Abundant small non-coding RNAs retained in spermatozoa include miRNAs and piRNAs. Some, like miR-34c are essential to the early embryo development required for the first cellular division. Others like the piRNAs are likely part of the genomic dance of confrontation and consolidation. Other non-coding spermatozoal RNAs include transposable elements, annotated lnc-RNAs, intronic retained elements, exonic elements, chromatin-associated RNAs, small-nuclear ILF3/NF30 associated RNAs, quiescent RNAs, mse-tRNAs and YRNAs. Some non-coding RNAs are known to act as epigenetic modifiers, inducing histone modifications and DNA methylation, perhaps playing a role in transgenerational epigenetic inherence. Transcript profiling holds considerable potential for the discovery of fertility biomarkers for both agriculture and human medicine. Comparing the differential RNA profiles of infertile and fertile individuals as well as assessing species similarities, should resolve the regulatory pathways contributing to male factor infertility. conclusions: Dad delivers a complex population of RNAs to the oocyte at fertilization that likely influences fertilization, embryo development, the phenotype of the offspring and possibly future generations. Development is continuing on the use of spermatozoal RNA profiles as phenotypic markers of male factor status for use as clinical diagnostics of the father's contribution to the birth of a healthy child. © The Author 2013. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.

Ambarish V.,Ms Ramaiah Medical College | Chandrashekara S.,Chanre Rheumatology and Immunology Center | Suresh K.P.,Indian National Institute of Animal Nutrition and Physiology
Indian Journal of Physiology and Pharmacology | Year: 2012

Exercises induce pro-inflammatory cytokines. We assessed the effect of different grades of exercises on inflammatory cytokine response. Twenty healthy volunteers performed a single bout of moderate exercise, a single bout of strenuous exercise and one month regular moderate exercise using standardized 10m Shuttle Walk Test. Interleukin-6 (IL-6) and Tumour Necrosis Factor Alpha (TNF-∝) were estimated by Sandwich ELISA method after each exercise regime. Statistics were run using SPSS software version 11.0, Systat software. Repeated measures ANOVA has been used for analysis of IL-6 values and Friedman test has been used for analyzing TNF-∝ and IL-6 values. Twenty healthy volunteers (18 to 30 years) were chosen for this study. The mean and SEM of plasma levels (pg/ ml) of IL-6 before exercise was 10.70±1.11 pg/ml, whereas, after acute moderate exercise and acute strenuous exercise it was 12.00±1.09 pg/ml and 13.35±0.89 pg/ml respectively. Interestingly, after one month of moderate exercise the values decreased to; 8.80±0.65 pg/ml. Mean and SEM of TNF-α before exercise was 121.78±29.06 pg/ml. With acute moderate exercise and after acute strenuous exercise the values were 132.90±35.75 pg/ml and 112.05±29.89 pg/ml respectively. After one month moderate exercise the levels decreased to 94.95±27.29 pg/ml. The observed changes in both IL-6 and TNF-α levels before and after both moderate and strenuous exercise were statistically significant. Although there was a slight decrease in the value of both the cytokines after one month of regular moderate exercise compared to baseline value, the difference in the values was not statistically significant. However, both IL-6 and TNF- levels showed overall statistically significant difference among the different grades of exercise. Plasma IL-6 and TNF- increase with acute moderate exercise and IL-6 increases further with acute strenuous exercise. Their levels tend to fall below baseline with one month of regular moderate exercise indicating that regular moderate exercise has beneficial effects.

Gurupriya V.S.,Indian National Institute of Animal Nutrition and Physiology | Divyashree B.C.,Indian National Institute of Animal Nutrition and Physiology | Roy S.C.,Indian National Institute of Animal Nutrition and Physiology
Theriogenology | Year: 2014

The postthaw motility and fertility of buffalo and cattle semen is reduced when they are cryopreserved for artificial insemination. In the present study, an attempt was made to characterize the cryogenic changes in proteases and antiprotease activities (APA) of buffalo and cattle semen because these proteolysis regulators have been reported to be associated with sperm motility and fertility. Buffalo sperm demonstrated at least two major proteases of 45 and 42 kDa and three minor proteases of 95, 52, and 33 kDa. Similarly, cattle sperm demonstrated three major proteases of 62, 45, and 42 kDa and two minor proteases of 85 and 78 kDa. Buffalo seminal plasma demonstrated at least three major proteases of 78, 68, and 62 kDa and one minor protease of 98 kDa and cattle seminal plasma demonstrated one major protease of 68 kDa and two minor proteases of 78 and 75 kDa. Except for the 45 kDa protease, most of the previously mentioned proteases were found to be metalloproteinases. Compared with fresh sperm, cryopreserved buffalo and cattle sperm demonstrated a major protease band of 52/49 kDa and the activity of this protease reduced progressively with the duration of cryopreservation. On the contrary, compared with the fresh seminal plasma, cryopreserved buffalo and cattle semen extenders displayed the presence of a new protease band of 45 kDa and demonstrated that this protease activity was leaked from buffalo and cattle cryopreserved spermatozoa. Buffalo and cattle seminal plasmas displayed at least two major APA of 86 and 26 kDa. Compared with buffalo, cattle seminal plasma demonstrated significantly greater APA. Thus, the present study demonstrated the presence of an array of proteases and APA in buffalo and cattle semen and the activities of which changed during cryopreservation. The leakage of the specific protease activity and changes in the proteases and APA might be attributed to reduced motility and fertility of cryopreserved semen in these species. © 2014 Elsevier Inc.

Roy S.C.,Indian National Institute of Animal Nutrition and Physiology | Ghosh J.,Indian National Institute of Animal Nutrition and Physiology
Molecular Reproduction and Development | Year: 2010

In ruminants, the phenomenon of endometrial tissue remodeling during the estrous cycle and early pregnancy is not fully understood. In this report, the occurrence of tissue remodeling, if any, in buffalo endometrium was studied by detecting gelatinases, matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs); the key regulators of tissue remodeling, in uterine luminal fluids (ULF) of cycling and early pregnant (approx. 43-65 days) buffaloes. Each stage of the estrous cycle and pregnant ULF demonstrated a unique profile of gelatinase activities compared to serum/follicular fluid, with a major gelatinase band of 60 kDa with highest activity in early-luteal stage. In addition to a 32 kDa uterus-specific gelatinase band detected in both non-pregnant and pregnant ULFs, the pregnant ULF displayed three new gelatinase bands of 86, 78, and 57 kDa. Western blot technique confirmed the presence of MMP-2 (54 kDa), MMP-9 (76/73 kDa), TIMP-1 (32 kDa), TIMP-2(20 kDa), and two molecular weight forms (31 and 22 kDa) of TIMP-3 in buffalo ULF with varying band intensities. Highest MMP-2 and MMP-9 activities were observed in follicular and early-luteal stage ULFs, respectively. Highest TIMP-1 activity was observed in early-luteal ULF. Interestingly, TIMP-2 activity was only detected in mid-luteal, late-luteal, and follicular stage ULFs with significantly increasing intensities. Highest activities of 31 and 22 kDa TIMP-3 were associated with late-luteal and early-luteal stage ULFs, respectively. The varied activities of MMPs and TIMPs in buffalo ULF during the estrous cycle and early pregnancy might be a reflection of dynamic structural remodeling of the endometrium and/or developing conceptus. © 2010 Wiley-Liss, Inc.

Suresh K.,Indian National Institute of Animal Nutrition and Physiology | Chandrashekara S.,ChanRe Rheumatology and Immunology Center and Research
Journal of Human Reproductive Sciences | Year: 2012

Determining the optimal sample size for a study assures an adequate power to detect statistical significance. Hence, it is a critical step in the design of a planned research protocol. Using too many participants in a study is expensive and exposes more number of subjects to procedure. Similarly, if study is underpowered, it will be statistically inconclusive and may make the whole protocol a failure. This paper covers the essentials in calculating power and sample size for a variety of applied study designs. Sample size computation for single group mean, survey type of studies, 2 group studies based on means and proportions or rates, correlation studies and for case-control for assessing the categorical outcome are presented in detail.

Gupta P.S.P.,Indian National Institute of Animal Nutrition and Physiology | Nandi S.,Indian National Institute of Animal Nutrition and Physiology
Reproduction in Domestic Animals | Year: 2010

Contents: The present study was undertaken to isolate buffalo preantral follicles (PFs), to test the viability and sizes of buffalo PFs and to examine the effect of various growth factors (insulin-like growth factor, fibroblast growth factor) and an antioxidant (β mercaptoethanol) on the in vitro growth, survival and antrum formation rates of buffalo PFs and growth rates of oocytes in cultured PFs. Preantral follicles from slaughtered buffalo ovaries were recovered by a combined mechanical and enzymatic method. The recovery rates of >40-100, 101-200, 201-300, 301-400 and 401-500 μm PFs were 5.1, 3.2, 3.1, 6.3 and 5.1 per ovary, respectively. The corresponding viability rates were 76.1%, 78.1%, 85.2%, 92.5% and 92.6%, respectively. There was a positive correlation (r = 0.73) between oocyte size and the follicular size. However, there was no significant correlation between the size of oocyte and its viability at the time of its retrieval from ovary. Insulin-like growth factor and fibroblast growth factor improved the survival of buffalo PFs and regulated their growth in culture. The growth factors and β mercaptoethanol in association synergically improved the growth and survival of buffalo PFs. © 2008 The Authors. Journal compilation. © 2008 Blackwell Verlag.

Samanta A.K.,Indian National Institute of Animal Nutrition and Physiology | Jayapal N.,Indian National Institute of Animal Nutrition and Physiology | Kolte A.P.,Indian National Institute of Animal Nutrition and Physiology | Senani S.,Indian National Institute of Animal Nutrition and Physiology | And 3 more authors.
Bioresource Technology | Year: 2012

In this study, a process for producing XOS from . Sehima nervosum grass was developed. The grass contains 28.1% hemicellulose. NaOH and steam application yielded 98% of original xylan in contrast to 85% by KOH application. Hydrolysis of xylan with commercial xylanase caused breakdown into XOS comprising of xylobiose, xylotriose along with xylose. Response surface model (RSM) revealed highest xylobiose yield (11. g/100. g xylan) at pH 5.03, temperature 45.19. °C, reaction time 10.11. h with enzyme dose 17.41. U. Similarly for maximizing xylotriose yield, ideal hydrolysis conditions were pH 5.11, temperature 40.33. °C, reaction time 16.55. h with enzyme dose 13.20. U. A two step process encompassing xylan fractionation and enzymatic hydrolysis enabled XOS production from the . S. nervosum grass. © 2012 Elsevier Ltd.

Suresh K.,Indian National Institute of Animal Nutrition and Physiology
Journal of Human Reproductive Sciences | Year: 2011

Randomization as a method of experimental control has been extensively used in human clinical trials and other biological experiments. It prevents the selection bias and insures against the accidental bias. It produces the comparable groups and eliminates the source of bias in treatment assignments. Finally, it permits the use of probability theory to express the likelihood of chance as a source for the difference of end outcome. This paper discusses the different methods of randomization and use of online statistical computing web programming (www.graphpad.com/quickcalcs or www.randomization.com) to generate the randomization schedule. Issues related to randomization are also discussed in this paper.

Bhatta R.,Indian National Institute of Animal Nutrition and Physiology | Saravanan M.,Indian National Institute of Animal Nutrition and Physiology | Baruah L.,Indian National Institute of Animal Nutrition and Physiology | Sampath K.T.,Indian National Institute of Animal Nutrition and Physiology
Journal of the Science of Food and Agriculture | Year: 2012

Background: Plant tannins as rumen modifiers are better than chemicals or antibiotic-based modifiers since these compounds are natural products which are environmentally friendly and therefore have a better acceptance with regard to feed safety issues. Tropical plants containing phenols such as tannins were found to suppress or eliminate protozoa from the rumen and reduce methane and ammonia production. The screening of these plants is an important step in the identification of new compounds and feed additives which might contribute to mitigate rumen methanogenesis. The present study was carried out to determine the efficacy of tannins from tropical tree leaves for their methane reduction properties. Results: Activity of tannins, as represented by the increase in gas volume with the addition of polyethylene glycol (PEG)-6000 as a tannin binder (tannin bioassay) was highest in Ficus bengalensis (555%), followed by Azardirachta indica (78.5%). PEG addition did not alter (P > 0.05) methane percentage in Ficus racemosa, Glyricidia maculata, Leucena leucocephala, Morus alba and Semaroba glauca, confirming that tannins in these samples did not affect methanogenesis. The increase (P < 0.05) in protozoa population with PEG was maximal in Ficus religiosa (50), followed by Moringa oleifera (31.2), Azardirachta indica (29.9) and Semaroba glauca (27.5). There was no change (P > 0.05) in the protozoa population in Autocarpus integrifolia, Ficus bengalensis, Jatropha curcus, Morus alba and Sesbania grandiflora, demonstrating that methane reduction observed in these samples per se was not due to defaunation effect of the tannin. The increase in total volatile fatty acid concentration in samples with PEG ranged from 0.6% to > 70%. The highest increase (%) in NH3-N was recorded in Azardirachta indica (67.4), followed by Ficus mysoriensis (35.7) and Semaroba glauca (32.6) leaves, reflecting strong protein binding properties of tannin. Conclusion: The results of our study established that in vitro methanogenesis was not essentially related to the density of protozoa population. Tropical tree leaves containing tannins such as Autocarpus integrifolia, Jatropha curcus and Sesbania grandiflora have the potential to suppress methanogenesis. Therefore tannins contained in these plants could be of interest in the development of new additives in ruminant nutrition. © 2012 Society of Chemical Industry.

Gupta P.S.P.,Indian National Institute of Animal Nutrition and Physiology | Nandi S.,Indian National Institute of Animal Nutrition and Physiology
Reproduction in Domestic Animals | Year: 2012

The development of efficient ovarian preantral follicle (PF) isolation and culture systems provide a large number of oocytes for the manipulation and embryo production. It also helps for understanding the mechanisms of follicle and oocyte development. Isolation and culture protocols for PFs were developed for many domestic species like cattle, buffalo, sheep, goat, pig, horse, camel, dog and cats; however, embryo production from oocytes derived from in vitro grown PFs was reported only in pigs, buffalo, sheep and goat. The rate of oocyte maturation from PFs grown in vitro is low and requires considerable research. This paper presents an overview of isolation and culture systems of PFs that have been developed for domestic species (cattle, buffalo, sheep, goat, pigs, horse, camel, dog and cat) along with the current status of progress achieved in the direction of producing embryos using PFs as the source of oocyte in these species. © 2011 Blackwell Verlag GmbH.

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